Thermo Fischer Scientific
CD25 Monoclonal Antibody (PC61.5), PerCP-Cyanine5.5, eBioscience™
45-0251-82
PC61.5
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In different healthy and pathological contexts, CD4 T cells differentiate into a range of specialized subsets, including FOXP3+ regulatory T cells expressing different levels of CD25 and transcription factor Helios. In mouse models of aging and autoimmune diseases we study the plasticity of effector and regulatory CD4 T cells. We used anti-CD25 to analyze the CD4+FOXP3+ T cell population in healthy control BALB/C mice.
Flow Cytometry
Splenocytes of wild-type 6 months-old BALB/C mouse
Washed twice in PBS and incubated with L/D fixable aqua dye 20 min at 4°C, then washed twice in PBS.
2.4G2
Surface staining with anti-CD3 BUV805, anti-CD4 BUV495, anti-CD25 PerCP Cy5.5 at 1/50, etc., in Cell Staining Buffer (Biolegend) incubated 30 min at 4°C.
Cells are then fixed and permeabilized with commercial kit (True Nuclear, BioLegend) 45 min at 4°C with fixative, and 15 min at 4°C with permeabilization buffer) and stained for transcription factor.
Blue laser on Symphony cytometer
The staining identifies different FOXP3+ subpopulation that express different levels of CD25 and Helios.
N/A
Visible, even after fixation and permeabilization.
Not very bright.
Good option, resist to permeabilization and fixation, but downregulated on CD4 T cells upon PMA/ionomycin restimulation.